PROJECT SUMMARY ? TR&D2 Drosophila serves as a model not only of human biology but also of the biology of other dipterans, including mosquito vectors of disease, and provides an instructive example of technologies that can be applied to direct study of other insects. In this project, we focus on development of functional genomics and proteomics resources for the study of mosquito vectors of disease, with a particular focus on development of CRISPR pooled screens for Aedes mosquitoes, which are vectors of viral pathogens including Zika, Dengue, and Chikungunya viruses, and for Anopheles mosquitoes, the vector for malaria. We will additionally develop a pipeline for isolation of robust protein binders, i.e. nanobodies, for use in visualization and in vivo functional blocking, based on a yeast nanobody display platform. A set of collaborators with established assays in mosquitoes will help us to test and improve the screening and nanobody technologies. Altogether, this project will provide much-needed reagents, including a platform for genome-wide screening, for direct study of pathogen entry and other aspects of mosquito biology relevant to infectious diseases and mosquito population control.